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High-performance liquid chromatography of transfer ribonucleic acids on spherical hydroxyapatite beads. II. Effects
Y Yamakawa1, K Miyasaka, T Ishikawa
1Department of Applied Immunology, National Institute of Health, Tokyo, Japan.
Abstract:
The effects of pH and sodium chloride on the high-performance liquid chromatography of transfer ribonucleic acids (tRNAs) on spherical hydroxyapatite beads were investigated. Binding of tRNAs on the beads was strengthened by increasing the pH of the mobile phase (phosphate buffer, pH 6.4-8.0), a phenomenon which is opposite to the retention of proteins on the beads. By using phosphate buffer of pH 8.0 instead of pH 6.8, the resolution of tRNAs was improved significantly; as many as ten times more theoretical plates were calculated with the use of the former buffer. Addition of sodium chloride to the phosphate buffer has a bifunctional effect on the retention of tRNAs: elution of tRNA(f-Met) and tRNA(Val) was retarded whereas that of tRNA(Phe) was facilitated.